2. What is plant tissue culture?
Plant tissue culture is a technique of growing plant
cells, tissues, organs, seeds or other plant parts in a
sterile environment on a nutrient medium
3. WHY?
• The production of clones of plants that
produce particularly good flowers, fruits, or
have other desirable traits.
• To quickly produce mature plants.
• The production of multiples of plants in the
absence of seeds or necessary pollinators to
produce seeds.
• The regeneration of whole plants from plant
cells that have been genetically modified.
4. WHY?
• The production of plants in sterile
containers reduces disease transmission
• Allows production of plants from seeds
that otherwise have very low chances
of germinating and growing,
i.e.: orchids and Nepenthes.
• To clean particular plants of viral and other
infections and to quickly multiply these
plants as 'cleaned stock'
for horticulture and agriculture.
5. Terminology
• Explant
– Living tissue transferred
from a plant to an artificial
medium for culture.
– It can be any portion of
the shoot, leaves, roots,
flower or cells from a plant.
6. How?
Adult plant cells are totipotent, meaning they have the ability to give rise to a fully
differentiated plant. Because of this, it is possible to collect cells from a mature plant and
use those cells to produce clones of that plant.
7. Plant tissue Culture Basics
• Modern plant tissue culture is performed under
aseptic conditions
• Living plant materials from the environment are
naturally contaminated on their surfaces (and
sometimes interiors) with microorganisms, so
surface sterilization of starting material (explants)
in chemical solutions (usually alcohol and sodium
or calcium hypochlorite is required).
8. Plant tissue Culture Basics
• Explants are then usually placed on the
surface of a solid culture medium, but are
sometimes placed directly into a liquid
medium, when cell suspension cultures are
desired.
• Culture media are generally composed of
inorganic salts plus a few organic nutrients,
vitamins and plant hormones.
12. Macronutrient salts
• Nitrogen – Influences plant growth rate, essential in
plant nucleic acids (DNA), proteins, chlorophyll, amino
acids, and hormones.
• Phosphorus – Abundant in meristematic and fast
growing tissue, essential in photosynthesis, respiration.
• Potassium – Necessary for cell division, meristematic
tissue, helps in the pathways for carbohydrate, protein
and chlorophyll synthesis.
13. Macronutrient salts
• Calcium - Involved in formation of cell walls and root and
leaf development. Participates in translocation of sugars,
amino acids, and ties up oxalic acid (toxin).
• Iron - Involved in respiration , chlorophyll synthesis and
photosynthesis. FeNaEDTA = sodium salt of EDTA
sequesters iron, making it available to plants.
• Magnesium - Involved in photosynthetic and respiration
systems. Active in uptake of phosphate and translocation
of phosphate and starches.
14. Micronutrient salts
• Sulfur - Involved in formation of nodules and chlorophyll
synthesis, structural component of amino acids and
enzymes.
• Manganese - Involved in regulation of enzymes and
growth hormones. Assists in photosynthesis and
respiration.
15. Micronutrient salts
• Molybdenum - Involved in enzymatic reduction of
nitrates to ammonia. Assists in conversion of inorganic
phosphate to organic form.
• Zinc - Involved in production of growth hormones and
chlorophyll. Active in respiration and carbohydrate
synthesis.
• Boron - Involved in production of growth hormones and
chlorophyll. Active in respiration and carbohydrate
synthesis.
• Copper -Involved in photosynthetic and respiration
systems. Assists chlorophyll synthesis and used as reaction
catalyst.
16. Organic Compounds
• Carbon Sources – Sucrose, sometimes
Glucose or Fructose (Plants Need Carbon)
• Vitamins –
– Adenine – part of RNA and DNA
– Inositol – part of the B complex, in phosphate form is
part of cell membranes, organelles and is not essential
to growth but beneficial
– Thiamine – essential as a coenzyme in the citric acid
cycle.
17. Still other organics
• Organic Acids
– Citric acid (150 mg/l) typically used with ascorbic
acid (100 mg/l) as an antioxidant.
– Can also use some of Kreb Cycle acids
• Phenolic compounds
– Phloroglucinol - Stimulates rooting of shoot
sections
18. Natural Complexes
• Coconut endosperm
• Fish emulsion
• Protein hydrolysates
• Tomato juice
• Yeast extracts
• Malt extract
• Potato agar
22. Gibberellin
• Not generally used in tissue culture
• Tends to suppress root formation and
adventitious embryo formation
23. Abscisic Acid
Primarily a growth inhibitor but enables more
normal development of embryos, both
zygotic and adventitious
24. Ethylene
• Question is not how much to add but how
to get rid of it in-vitro
• Natural substance produced by tissue
cultures at fairly high levels especially when
cells are under stress
• Enhances senescense
• Supresses embryogenesis and development
in general
25. Hormone Combinations
• Callus development
• Adventitious embryogenesis
• Rooting of shoot cuttings
• Adventitious shoot and root formation
26.
27. Applications[edit]
Plant tissue culture is used widely in the plant sciences, forestry, and in horticulture.
Applications include:
The commercial production of plants used as potting, landscape, and florist subjects,
which uses meristem and shoot culture to produce large numbers of identical
individuals.
1.To conserve rare or endangered plant species.[6]
2.A plant breeder may use tissue culture to screen cells rather than plants for
advantageous characters, e.g. herbicide resistance/tolerance.
3.Large-scale growth of plant cells in liquid culture in bioreactors for production of
valuable compounds, like plant-derived secondary metabolites and recombinant
proteins used as biopharmaceuticals.[7]
4.To cross distantly related species by protoplast fusion and regeneration of the
novel hybrid.
]
28. 5.To rapidly study the molecular basis for physiological, biochemical, and reproductive
mechanisms in plants, for example in vitro selection for stress tolerant plants.[8]
6.To cross-pollinate distantly related species and then tissue culture the resulting embryo
which would otherwise normally die (Embryo Rescue).
7.For chromosome doubling and induction of polyploidy,[9] for example doubled
haploids, tetraploids, and other forms of polyploids. This is usually achieved by application
of antimitotic agents such as colchicine or oryzalin.
8.As a tissue for transformation, followed by either short-term testing of genetic constructs
or regeneration of transgenic plants.
9.Certain techniques such as meristem tip culture can be used to produce clean plant
material from virused stock, such as sugarcane[10], potatoes and many species of soft fruit.
Production of identical sterile hybrid species can be obtained.
10.Large scale production of artificial seeds through somatic embryogenesis[11
